SANGUINARINE IS REDUCED BY NADH THROUGH A COVALENT ADDUCT

SÁNDOR, Roman, Jiří SLANINA, Adam MIDLIK, Kristýna ŠEBRLOVÁ, Lucie NOVOTNÁ, Petr TÁBORSKÝ, Eva TÁBORSKÁ and Ondřej PEŠ. SANGUINARINE IS REDUCED BY NADH THROUGH A COVALENT ADDUCT. In 20. ročník Školy hmotnostní spektrometrie. 2019. ISBN 978-80-88195-14-6.

Basic information

• Original name: SANGUINARINE IS REDUCED BY NADH THROUGH A COVALENT ADDUCT
• Authors: SÁNDOR, Roman (703 Slovakia, belonging to the institution), Jiří SLANINA (203 Czech Republic, belonging to the institution), Adam MIDLIK (703 Slovakia, belonging to the institution), Kristýna ŠEBRLOVÁ (203 Czech Republic, belonging to the institution), Lucie NOVOTNÁ (203 Czech Republic, belonging to the institution), Petr TÁBORSKÝ (203 Czech Republic, belonging to the institution), Eva TÁBORSKÁ (203 Czech Republic, belonging to the institution) and Ondřej PEŠ (203 Czech Republic, guarantor, belonging to the institution).
• Edition: 20. ročník Školy hmotnostní spektrometrie, 2019.

Other information

• Original language: English
• Type of outcome: Conference abstract
• Field of Study: 30104 Pharmacology and pharmacy
• Country of publisher: Czech Republic
• Confidentiality degree: is not subject to a state or trade secret
• WWW: URL
• RIV identification code: RIV/00216224:14110/19:00110706
• Organization unit: Faculty of Medicine
• ISBN: 978-80-88195-14-6
• Keywords in English: benzophenanthridine alkaloids; metabolism; LC MS; NADH; biotransformation; ene cyclization
• Tags: rivok

Abstract

Benzo[c]phenanthridine alkaloids are a family of compounds with cytotoxic, anti-fungal and antimicrobial properties, which are heavily metabolized by the CYP 450 system. Most of the research of these compounds has been focused rather on their biological effects than their metabolism; however, it has already been found that the first step of their biotransformation is a reduction to their respective dihydro forms. Interestingly, this reaction step may proceed with NAD(P)H even when no additional enzymatic apparatus is present. We decided to study this interaction with sanguinarine (SA), as this effect has been noted but never examined any further. From the results of Matkar et al.[1], who found an unexpected compound on RP-TLC and 1D electrophoresis of NADH and SA, we hypothesized that SA forms a negatively charged, covalent adduct with NADH. We used TLC and LC-MS to identify the compound, observed at 997 Da (singly charged) and 499 Da (doubly charged). The kinetics of the equimolar mixture and the kinetics of the adduct isolated by semi-preparative chromatography was studied by the UV-Vis and MS/MS-MS spectrometry to infer the state of all reaction compounds at different time points. The kinetics of the equimolar mixture showed a decrease in concentration of SA while the concentration of the reduced form, dihydrosanguinarine, and the adduct were increasing. The kinetics of the isolated adduct was more complicated, as the adduct partially decomposed to the reactants. The results strongly supported the hypothesis that during the non-enzymatic reduction of SA by NADH, a covalent semi-stable adduct was present, and its decomposition to products was the rate-limiting step.

Links

• MUNI/A/0976/2018, interní kód MU: Name: Příspěvek chemických a biochemických metodik ke studiu molekulární podstaty vybraných patologických stavů a onemocnění

Investor: Masaryk University, Category A

• MUNI/A/1550/2018, interní kód MU: Name: Farmakologický výzkum v oblasti farmakokinetiky, neuropsychofarmakologie a onkologie

Investor: Masaryk University, Category A